Glycyrrhizin and its derivatives promote hepatic differentiation via sweet receptor, Wnt, and Notch signaling

Akihiro Morita; Yuta Omoya; Rie Ito; Yuya Ishibashi; Keiichi Hiramoto; Shiho Ohnishi; Nobuji Yoshikawa; Shosuke Kawanishi

doi:10.1016/j.bbrep.2021.101181

Glycyrrhizin and its derivatives promote hepatic differentiation via sweet receptor, Wnt, and Notch signaling

Biochem Biophys Rep. 2021 Dec 4:28:101181. doi: 10.1016/j.bbrep.2021.101181. eCollection 2021 Dec.

Authors

Akihiro Morita¹, Yuta Omoya¹, Rie Ito¹, Yuya Ishibashi¹, Keiichi Hiramoto¹, Shiho Ohnishi¹, Nobuji Yoshikawa², Shosuke Kawanishi¹

Affiliations

¹ Department of Pharmaceutical Sciences, Suzuka University of Medical Science, Suzuka, Mie, 513-8670, Japan.
² Matsusaka R&D Center, Cokey Co., Ltd., Matsusaka, Mie, 515-0041, Japan.

Abstract

The acute liver disease is involved in aberrant release of high-mobility group box 1 (HMGB1). Glycyrrhizin (GL), a traditional Chinese medicine for liver disease, binds to HMGB1, thereby inhibits tissue injury. However the mode of action of GL for chronic liver disease remains unclear. We investigated the effects of glycyrrhizin (GL) and its derivatives on liver differentiation using human iPS cells by using a flow cytometric analysis. GL promoted hepatic differentiation at the hepatoblast formation stage. The GL derivatives, 3-O-mono-glucuronyl 18β-glycyrrhetinic acid (Mono) and 3-O-[glucosyl (1 → 2)-glucuronyl] 18β-glycyrrhetinic acid increased AFP⁺ cell counts and albumin⁺ cell counts. Glucuronate conjugation seemed to be a requirement for hepatic differentiation. Mono exhibited the most significant hepatic differentiation effect. We evaluated the effects of (±)-2-(2,4-dichlorophenoxy) propionic acid (DP), a T1R3 antagonist, and sucralose, a T1R3 agonist, on hepatic differentiation, and found that DP suppressed Mono-induced hepatic differentiation, while sucralose promoted hepatic differentiation. Thus, GL promoted hepatic differentiation via T1R3 signaling. In addition, Mono increased β-catenin⁺ cell count and decreased Hes5⁺ cell count suggesting the involvement of Wnt and Notch signaling in GL-induced hepatic differentiation. In conclusion, GL exerted a hepatic differentiation effect via sweet receptor (T1R3), canonical Wnt, and Notch signaling.

Keywords: AFP, α-fetoprotein; Api, 3-O-[apiosyl (1 → 2)-glucuronyl] βGA; CBX, carbenoxolone, 3-O-hemisuccinyl βGA; CK-19, cytokeratin 19; DMSO, dimethyl sulfoxide; DP, (±)-2-(2,4-dichlorophenoxy) propionic acid; GL, glycyrrhizin; Glc, 3-O-[glucosyl (1 → 2)-glucuronyl] βGA; Glycyrrhizin; HMGB1, high-mobility group box1; HNF-4α, hepatocyte nuclear factor 4α; Hepatic differentiation; Hes, hairy and enhancer of split; LSG, licorice saponin G; LSH, licorice saponin H; Liver regeneration; Mono, 3-O-mono-glucuronyl βGA; Sweet receptor; T1R3; αGA, 18α-glycyrrhetinic acid; βGA, 18β-glycyrrhetinic acid.